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Open Access Research article

A novel quadruplex real-time PCR method for simultaneous detection of Cry2Ae and two genetically modified cotton events (GHB119 and T304-40)

Xiang Li1*, Xiuxiu Wang2, Jielin Yang1, Yueming Liu1, Yuping He1* and Liangwen Pan1

Author Affiliations

1 GMO Detection Laboratory, Shanghai Entry-Exit Inspection and Quarantine Bureau, 1208 Minsheng Road, Shanghai 200135, PR China

2 Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, PR China

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BMC Biotechnology 2014, 14:43  doi:10.1186/1472-6750-14-43

Published: 16 May 2014

Abstract

Background

To date, over 150 genetically modified (GM) crops are widely cultivated. To comply with regulations developed for genetically modified organisms (GMOs), including labeling policies, many detection methods for GMO identification and quantification have been developed.

Results

To detect the entrance and exit of unauthorized GM crop events in China, we developed a novel quadruplex real-time PCR method for simultaneous detection and quantification of GM cotton events GHB119 and T304-40 in cotton-derived products (based on the 5′-flanking sequence) and the insect-resistance gene Cry2Ae. The limit of detection was 10 copies for GHB119 and Cry2Ae and 25 copies for T304-40. The limit of quantification was 25 copies for GHB119 and Cry2Ae and 50 copies for T304-40. Moreover, low bias and acceptable standard deviation and relative standard deviation values were obtained in quantification analysis of six blind samples containing different GHB119 and T304-40 ingredients.

Conclusions

The developed quadruplex quantitative method could be used for quantitative detection of two GM cotton events (GHB119 and T304-40) and Cry2Ae gene ingredient in cotton derived products.

Keywords:
Cry2Ae; Event-specific; GHB119; Quadruplex real-time PCR; T304-40